Bioengineering a Chimeric Human Lung

描述（由申请人提供）：美国有近2500万人患有终末期肺病。肺移植是这些患者唯一的治疗选择，但仍然受到供体器官短缺、长期排斥和免疫抑制治疗需求的阻碍。制造含有患者自身细胞的肺的能力将从根本上改变我们目前治疗终末期肺病的方式。我们建议用人类多能干细胞（hPSC）衍生的肺祖细胞播种部分去细胞化的供体肺，以实现嵌合人肺的生物工程。我们的目标是：(1)获得hPSC规范对肺谱系和使用天然肺基质形成肺组织的新见解；(2)生物工程功能肺用于移植。我们设想，排斥/边缘质量供体肺的功能可以通过患者的hpsc来源的肺细胞部分替代细胞材料来改善。我们的假设是，植入肺去细胞化区域的hpsc祖细胞将被诱导再生和重塑供体肺，以响应来自组织基质和残余细胞的部位特异性信号。我们的方法是将被拒绝移植的供体肺从肺的有限区域移除细胞，同时保留脱细胞基质的组成、结构和机械特性以及周围完整的实质。同时通过肺实质灌注脱细胞液和通过门静脉灌注Perfadex溶液，我们将保持完整的肺血管系统。脱细胞区随后将被hpsc衍生的肺祖细胞重新填充，以产生能够在重塑后改善的最低可接受水平的气体交换的肺。目的1是从hPSCs中获得并充分表征肺和气道上皮的不同谱系。目的2：通过在脱细胞基质切片上培养hpsc衍生的肺细胞，对人肺组织进行生物工程。目的3是通过用hPSC衍生的肺细胞重新填充肺的脱细胞区域来形成嵌合人肺，并使用临床肺灌注系统研究功能恢复。该提案的最终目标是通过结合三个主要的创新组成部分来实现因移植而被拒绝的供肺的功能恢复：(1)从hPSCs中衍生肺细胞；(2)在保留脉管系统和实质结构的情况下对供肺进行区域脱细胞；(3)通过用受体细胞重新填充肺来实现功能恢复。

DESCRIPTION (provided by applicant): Nearly 25 million people in the United States suffer from end-stage lung disease. Lung transplantation, the only curative option for these patients, remains hampered by donor organ shortages, long-term rejection, and the need for immunosuppressive therapy. The ability to generate lungs containing a patient's own cells would radically change the way we currently treat end-stage lung disease. We propose to bioengineer chimeric human lungs by seeding partially decellularized donor lungs with human pluripotent stem cell (hPSC) derived pulmonary progenitors. Our goals are to (1) obtain new insights into hPSC specification towards pulmonary lineages and the formation of lung tissue using native lung matrix, and (2) bioengineer functional lungs for transplantation. We envision that the function of rejected/marginal quality donor lungs can be improved by partial replacement of cellular material by a patient's hPSC-derived pulmonary cells. Our hypothesis is that hPSC-progenitors seeded into the decellularized regions of the lung will be induced to regenerate and remodel the donor lung in response to site-specific signals from the tissue matrix and residual cells. Our approach will be to take donor lungs rejected for transplantation and remove cells from limited regions of the lung while preserving the composition, architecture, and mechanical properties of the decellularized matrix and the surrounding intact parenchyma. By simultaneously perfusing decellularization fluids through the lung parenchyma and Perfadex solution through the portal vein, we will preserve intact lung vasculature. The decellularized regions will then be repopulated by hPSC-derived lung progentor cells to produce a lung that is capable of some minimally acceptable level of gas exchange that will improve upon remodeling. Aim 1 is to derive and fully characterize the different lineages of the lung and airway epithelium from hPSCs. Aim 2 is to bioengineer human lung tissue by hPSC-derived pulmonary cells cultured on slices of decellularized matrix. Aim 3 is to form a chimeric human lung by repopulating decellularized regions of the lung with hPSC- derived pulmonary cells and investigate functional recovery using a clinical lung perfusion system. This proposal ultimately aims to achieve functional recovery of donor lungs rejected for transplantation by combining three major innovative components: (1) Derivation of pulmonary cells from the hPSCs, (2) Regional decellularization of the donor lung with the preservation of vasculature and parenchymal architecture, and (3) Functional recovery through repopulation of the lung with the recipient's cells.